Process for preparing a collagen-rich composition

ABSTRACT

This disclosure provides a high collagen composition prepared from a poultry broth and methods of making the same by filtration.

RELATED APPLICATIONS

This application is a continuation-in-part of U.S. patent applicationSer. No. 16/220,761 filed Dec. 14, 2018, now U.S. Pat. No. 10,694,768,which is a continuation of International Application No.PCT/US2017/037759 filed Jun. 15, 2017, which claims the benefit ofpriority to U.S. Patent Application No. 62/350,456 filed Jun. 15, 2016,U.S. Patent Application No. 62/461,058 filed Feb. 20, 2017, and U.S.Patent Application No. 62/461,061 filed Feb. 20, 2017. This applicationis also a continuation-in-part of U.S. patent application Ser. No.16/220,728 filed Dec. 14, 2018, now U.S. Pat. No. 10,694,767, which is acontinuation of International Application No. PCT/US2017/037756 filedJun. 15, 2017, which claims the benefit of priority to U.S. PatentApplication No. 62/350,456 filed Jun. 15, 2016, U.S. Patent ApplicationNo. 62/461,058 filed Feb. 20, 2017, and U.S. Patent Application No.62/461,061 filed Feb. 20, 2017. Both U.S. patent application Ser. No.16/220,761 and U.S. patent application Ser. No. 16/220,728 are alsocontinuation-in-part of U.S. patent application Ser. No. 14/698,332filed Apr. 28, 2015, which claims the benefit of priority to U.S.Provisional Application No. 61/985,252 filed Apr. 28, 2014. Both U.S.patent application Ser. No. 16/220,761 and U.S. patent application Ser.No. 16/220,728 are also continuations-in-part of U.S. patent applicationSer. No. 14/850,405 filed Sep. 10, 2015, now U.S. Pat. No. 10,349,669,which claims the benefit of priority to U.S. Provisional Application No.62/048,648 filed Sep. 10, 2014. The entire content of all of theabove-mentioned applications is hereby incorporated, by reference intothis application.

BACKGROUND 1. Field of the Invention

This disclosure relates to a process for making a broth composition.More particularly, the disclosure relates to a pumpable compositionprepared from poultry or other animal or plant sources.

2. Description of Related Art

Broth prepared from animals (including poultry) has high nutritionvalues. However, most concentrated broth compositions are not pumpable(or pourable) and may be difficult to handle and transport. Methods forpreparing pumpable broth compositions have been disclosed. See e.g.,U.S. patent application Ser. Nos. 14/210,284 and 14/850,405. However,existing methods typically require addition of enzymes or a rawextraction step.

Methods have also been disclosed for preparing collagen rich compositionfrom poultry products; however, most existing methods involve many stepsand typically require some chemical or enzymatic treatment step.

SUMMARY

The instrumentalities disclosed herein overcome the problems outlinedabove by providing a method of preparing a composition enriched withcollagen. In one embodiment, the composition is enriched with Type IIcollagen. In another embodiment, the enriched Type II collagen issoluble in water at room temperature.

In one embodiment, the disclosed process may include: (a) applying astarting material to a filtration means, said filtration means having apore size of 500 nanometers (nm) or smaller, said starting materialbeing prepared from an animal source; (b) allowing the startingcomposition to pass through the filtration means, and (c) collectingretentate that is retained on said filtration means to obtain saidcomposition, wherein said composition obtained in step (c) comprisesmore than 3%, 4%, or 5% hydroxyproline by weight of total amino acids insaid composition. In one aspect, retentate obtained in step (c)comprises more than 50%, 70%, 80% or 90% (w/w) collagen. In anotheraspect, the collagen comprises more than 50%, 70%, 80% or 90% (w/w) TypeII collagen. See Trentham et al., Science, 261:1727-30 (1993).

In one aspect, the starting material may be obtained by extractingproteins from poultry parts. In another aspect, the poultry partscontain 50-100% (w/w) of poultry bones and cartilage. In another aspect,the poultry parts contain 50-100%, 80-100%, or 90-100% (w/w) of poultrycartilage.

In another embodiment, the process may further include a cooking stepprior to step (a), wherein the poultry parts are cooked in water at atemperature 250 F or higher for 6 hours or longer during the cookingstep. In another embodiment, no enzyme is added to said startingmaterial before being applied to the filtration means. In anotherembodiment, the filtration means has a pore size of smaller than 500 nm,200 nm, 100 nm, or 50 nm.

In another embodiment, the process may include a step of collecting bothretentate that is retained on the filtration means and permeate thatpasses through said filtration means. The retentate is used as acollagen-rich ingredient while the permeate is used as a pumpablecomposition.

The present disclosure also provides a pumpable broth composition havinghigh percentage of solids and relatively long shelf life. In oneembodiment, no enzymes are used in the disclosed process. In anotherembodiment, no raw extraction step is used in the disclosed process. Thecomposition may be prepared from an animal or plant source. Examples ofanimal sources may include but are not limited to meat or other bodyparts of birds (e.g., poultry), cattle (beef), pigs (pork), amongothers. Examples of birds may include but are not limited to chickens orturkeys.

In one embodiment, the disclosed process for making a broth compositionthat is pumpable or pourable at refrigerated temperature may include (a)applying a starting composition to a filtration means, (b) allowing thestarting composition to pass through the filtration means, and (c)collecting permeate that passes through the filtration means to obtainthe pumpable composition. In another embodiment, the compositionobtained from step (c) has at least 50%, 60%, 70%, 80%, or 90% (w/w)solids. In another embodiment, the composition obtained from step (c)may be concentrated to obtain a pumpable composition that has at least50%, 60%, 70%, 80%, or 90% (w/w) solids.

In one embodiment, starting broth or extracts prepared from an animalsource (“starting composition” or “starting material”) are passedthrough a filtration means to separate the flow into permeates andretentates. In one aspect, the starting composition may be applied to afiltration means, and allowed to pass through the filtration means.Permeate that passes through said filtration means may then be collectedto obtain the pumpable composition. In one aspect, the filtration meansmay have a pore size of about 10 micrometers (μm), 5 μm, 2 μm, 500nanometers (nm), 250 nm, 100 nm, 90 nm, 80 nm, 70 nm, 60 nm, 50 nm, orsmaller. In another aspect, the filtration means may have pore sizebetween 300 nm and 1,000 nm, between 100 nm and 500 nm, between 300 nmand 500 nm, between 10 nm and 50 nm, or between 50 nm and 300 nm.

In one aspect, the pumpable composition obtained according to thedisclosed process may contain less collagen or collagen-derived proteinsthan the starting composition. For instance, it may contain less than10%, 5%, 3%, 2%, or 1% (w/w) of collagen. In another aspect, thestarting composition (broth or extract) may not be pumpable, but thepermeate is pumpable because significant amount of collagen orcollagen-derived proteins is retained by the filtration means. Inanother aspect, the permeate or retentate may have higher levels ofcertain essential amino acids (EAA)/indispensable amino acids (IAA) thanthe starting composition.

In another embodiment, the percentage of hydroxyproline by weight oftotal amino acids in the pumpable composition prepared according to thedisclosed processes is less than 5%, less than 4%, less than 3%, or lessthan 2% (w/w).

In another embodiment, the percentage of proline by weight of totalamino acids in the disclosed pumpable composition is less than 10%, lessthan 8%, less than 7%, or less than 5% (w/w).

In another embodiment, the percentage of glycine by weight of totalamino acids in the disclosed pumpable composition is less than 15%, lessthan 12%, less than 11%, or less than 10% (w/w).

In another embodiment, the filtration means may include are not limitedto microfiltration, ultrafiltration, nanofiltration, reverse osmosis,membrane or combination thereof. In another embodiment, the filtrationmeans may also include ion-exchange and elution, dialysis,centrifugation, or preparative gel filtration, among others.

In one aspect, this filtration means may specifically retain certainamino acids or compounds, resulting in permeate or retentate havinghigher concentration of these amino acids or compounds than the startingbroth or extracts. In one embodiment, the amino acids or compounds thatare enriched in either the permeate or retentate may include but are notlimited to collagen protein, peptides such as taurine, anserine,carnosine, creatine/creatinine, L-carnitine, choline, and other specificnutrients of value.

In another embodiment, the composition prepared according to thedisclosed process may have 0.85, 0.80, 0.75, 0.7, 0.6, 0.5 or lowerwater activity. In one aspect, the higher solids of the disclosedpumpable broth composition may help control microbial growth and preventspoilage of the broth products. In another aspect, the low wateractivity of the composition may help control microbial growth andprevent spoilage of the broth products. In one embodiment, the disclosedmay be store at ambient (room) temperature for extended time, forexample, for 10 days, 30 days, 60 days, 6 months, 12 months, or 24months, without microbial contamination. In another embodiment, becauseof the longer shelf life, the disclosed composition may be stored andshipped without requiring refrigeration.

In one embodiment, the starting composition may be any broth (extracts)prepared from an animal source. In another embodiment, one or moreenzymes may be used in the process of making the initial soluble proteincompositions. In another embodiment, no extraneous enzymes are used inthe process of preparing the starting composition or the final pumpablecomposition.

In one embodiment, salt(s) may be used in the disclosed process to helpprepare the pumpable composition. In another embodiment, no salt is usedin the disclosed process.

In another embodiment, the starting composition may be a suspension withinsolubles that may be separated by centrifugation before the startingcomposition is applied to the filtration means. In another embodiment,the starting composition may be prepared from a composition in solidform (e.g., powder) and made into a liquid form before being applied tothe filtration means.

The disclosed composition may be prepared from a starting materialderived from an animal source. For instance, the starting material maybe derived from chicken, turkey, beef, pork or other animal or poultrysources.

In one embodiment, the starting material may be prepared from rawmaterials. In another embodiment, the starting material may be preparedfrom previously cooked materials. The raw material may include but arenot limited to meat, trims, bones, skin, other animal parts orcombination thereof.

In another embodiment, the starting material may be in a substantiallyliquid form. The term “substantially liquid form” means that thestarting material is mostly liquid but may contain minor amount ofinsoluble material.

In another embodiment, the starting material may be obtained byextracting raw mechanically separated poultry (MSP), mechanicallyseparated chicken (MSC), or finely ground poultry pieces (such aspoultry trims or ground poultry parts) with water at room temperature orlower. By way of example, the extraction may be conducted by addingwater into raw MSC. The mixture can then be stirred to facilitate mixingand extracting. The ratio between the MSC and water in the extractionmixture may range from about 4:1 to about 1:20 by weight, from about 1:1to about 1:4 by weight, or about 1:2 by weight. In another embodiment,the MSC and water mixture may be subject to centrifugation at the end ofthe extraction. The liquid phase resulting from the centrifugation maybe collected and used as the starting material for preparing thepumpable broth composition of the present disclosure. In one aspect ofthis disclosure, the centrifugation may be performed at a speed of atleast 1,000 rpm, 2,000 rpm, or at least 3,500 rpm.

The starting material may be prepared on-site and may be used for makingthe present composition right after it is made fresh on-site.Alternatively, the starting material may be from packaged products ormay be collected off-site.

In another embodiment, the pumpable composition obtained in step (c) isfurther subject to a step (e) to remove sodium from the pumpablecomposition. In one aspect, the level of sodium may be reduced by atleast 30%, 50%, or 70% in step (e).

In another embodiment, the color of the pumpable composition obtained instep (c) is substantially lighter than the color of the startingcomposition. The color difference may be determined by human eyes or byan instrument, such as a spectrophotometer.

In another embodiment, the disclosed composition thus obtained may beused in numerous products. Examples of such products may include but arenot limited to protein drink, smoothies, sports drink or nutritionalbeverages.

DETAILED DESCRIPTION

This disclosure relates to a process for making a high proteincomposition from an animal source. In one aspect, the disclosed processmay be used to make a pumpable composition from a composition that isnot pumpable.

The term “refrigerated (or refrigeration) temperature” refers to atemperature ranging from about 0 C to about 10 C, for example, 4 C.

The term “pumpable” or pourable” refers to the state of a compositionhaving sufficient liquidity such that the composition may be poured bydecanting or it may be passed through a pump without applyingsignificant pressure. In one embodiment, the pumpable composition (e.g.,broth) has a viscosity of 50,000 cP, 25,000 cP, 10,000 cP, 5,000 cP orlower at 25 C.

In one embodiment, the disclosure provides methods of making a highquality protein composition from poultry. Poultry (e.g., chicken orturkey) is widely consumed in numerous applications as a healthy,nutritious food. Chicken broth is also widely used as the foundation formany classic foods including soups, stews, chowders, gravies, andsauces.

As compared to other broth products, the disclosed compositions areeasier to handle and provide a better balance of proteins and aminoacids than regular broth prepared according to conventional methods.

In one aspect, the disclosed process may be used to turn lower value rawpoultry materials into a high value protein powder or broth withoutusing additives.

By way of example, several embodiments of the disclosed processes aredescribed below:

1. In one embodiment, a method of making a pumpable composition isdisclosed, comprising:

-   -   (a) applying a starting composition to a filtration means, said        filtration means having a pore size of 500 nm or smaller,    -   (b) allowing the starting composition to pass through the        filtration means, and    -   (c) collecting permeate that passes through said filtration        means to obtain said pumpable composition,

wherein said composition obtained in step (c) is pumpable atrefrigeration temperature.

2. In another embodiment, methods according to Item 1 are disclosed,wherein the pumpable composition obtained in step (c) has at least 50%(w/w) solids.

3. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein said pumpable composition obtained in step(c) has collagen concentration of 10% (w/w) or lower.

4. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein hydroxyproline constitutes less than 3%(w/w) of total amino acids in said pumpable composition obtained in step(c).

5. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein proline constitutes less than 7% (w/w) oftotal amino acids in said pumpable composition obtained in step (c).

6. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein glycine constitutes less than 11% (w/w) oftotal amino acids in said pumpable composition obtained in step (c).

7. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein said filtration means is selected from thegroup consisting of microfiltration, ultrafiltration, nanofiltration,reverse osmosis, membrane and combination thereof.

8. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein the filtration means has a pore sizebetween 100 nm and 500 nm.

9. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein said animal source is poultry.

10. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein said starting composition is subjected to aseparation step (d) before being applied to the filtration means. In oneembodiment, step (d) may include, for example, phase separation ofliquid:liquid or liquid:solid in time with or without elevatedtemperature to obtain or siphon off the concentrated liquid or solid.

11. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein said separation step (d) is carried out bycentrifugation.

12. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein said pumpable composition obtained in step(c) has a water activity of 0.85 or lower.

13. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein enzyme is added to said startingcomposition before being applied to the filtration means.

14. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein salt is added to said starting compositionbefore being applied to the filtration means.

15. In another embodiment, methods according to any one of the precedingitems are disclosed, further comprising a step (e) to remove sodium fromthe pumpable composition obtained in step (c).

16. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein the level of sodium is reduced by at least50% in step (e).

17. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein the composition obtained in step (c) ispumpable at 4 C.

18. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein the color of the pumpable compositionobtained in step (c) is substantially different from the color of thestarting composition.

19. In another embodiment, methods according to any one of the precedingitems are disclosed, wherein the color of the pumpable compositionobtained in step (c) is substantially lighter than the color of thestarting composition.

In one embodiment, the soluble protein compositions may be used as aningredient in food or beverage products. In another embodiment, thedisclosed composition may also be used to prepare protein drinks,smoothies, or other nutritional or sport beverages.

It is to be noted that, as used in this specification and the claims,the singular forms “a,” “an,” and “the” include plural referents unlessthe context clearly dictates otherwise. Thus, for example, reference to“a device” may include reference to one device, as well as two or moredevices, unless the context clearly limits the reference to one device.

The terms “between” and “at least” as used herein are inclusive. Forexample, a range of “between 5 and 10” means any amount equal to orgreater than 5 but equal to or smaller than 10.

Unless otherwise specified, the percentage of certain component in acomposition is by weight of total solid. Various commercially availableproducts may have been described or used in this disclosure. It is to berecognized that these products are cited for purpose of illustrationonly. Certain physical and/or chemical properties and composition of theproducts may be modified without departing from the spirit of thepresent disclosure. One of ordinary skill in the art may appreciate thatunder certain circumstances, it may be more desirable or more convenientto alter the physical and/or chemical characteristics or composition ofone or more of these products in order to achieve the same or similarobjectives as taught by this disclosure.

EXAMPLES

The following examples are provided to illustrate the present invention,but are not intended to be limiting. The reagents, materials andinstruments are presented as typical components, and varioussubstitutions or modifications may be made in view of the foregoingdisclosure by one of skills in the art without departing from theprinciple and spirit of the present invention.

Example 1 Preparation of Pumpable Broth by Filtration

Frozen cooked chicken stock with 29% solids was received and thawed. Thestock was then diluted to 12.85% solids, which was passed through a 50nanometer ceramic membrane. The permeate was concentrated to 58% (w/w)solids. The concentrate had water activity measuring at 0.83. Thispermeate concentrate had high fluidity at refrigeration temperature(e.g., 4 C).

Amino analyses of the permeate and retentate show that hydroxyproline,proline, and glycine were much higher in the Retentate samples than inthe Permeate. This result confirmed that the majority of collagen andgelatin proteins was in the Retentate fraction while the percentage ofcollagen (and collagen derived proteins) and gelatin proteins in thePermeate fraction was reduced. This reduction of collagen and gelatinproteins in the Permeate fraction likely contributed to the reduction ingel strength and the increase in fluidity.

TABLE 1 Amino acid analyses of the permeate and retentate CalculatedCalculated W/W % to 100% W/W% to 100% Units Permeate solids Retentatesolids Hydroxyproline 0.27 1.16 0.83 4.07 Aspartic Acid 0.57 2.45 1.025.00 Threonine 0.27 1.16 0.40 1.96 Serine 0.29 1.24 0.42 2.06 GlutamicAcid 1.88 8.07 2.55 12.50 Proline 0.65 2.79 1.11 5.44 Glycine 1.02 4.381.89 9.26 Alanine 0.61 2.62 1.15 5.64 Cysteine 0.08 0.34 0.07 0.34Valine 0.25 1.07 0.43 2.11 Methionine 0.10 0.43 0.21 1.03 Isoleucine0.20 0.86 0.36 1.76 Leucine 0.36 1.55 0.79 3.87 Tyrosine 1.28 5.49 0.693.38 Phenylalanine 0.21 0.90 0.35 1.72 Hydroxylysine 0.04 0.17 0.10 0.49Ornithine 0.02 0.09 0.01 0.05 Lysine 0.55 2.36 1.05 5.15 Histidine 0.612.62 0.36 1.76 Arginine 0.41 1.76 0.93 4.56 Tryptophan 0.04 0.17 0.050.25 Total 9.71 14.77 72.40 Crude Protein* 17.77 76.27 18.28 89.61Potassium 1.72 7.38 0.717 3.51 Magnesium 0.028 0.12 0.014 0.07 Iron(ppm) 12.4 53.22 8.51 41.72 Zinc (ppm) 3.60 15.45 2.28 11.18 Calcium0.018 0.08 0.012 0.06 Sodium 0.775 3.33 0.342 1.68 Phosphorus 0.442 1.900.189 0.93

REFERENCES

All references listed below as well as publications, patents, patentapplications cited throughout this disclosure are hereby incorporatedexpressly into this disclosure as if fully reproduced herein.

-   Shah et al., U.S. patent application Ser. No. 10/912,560.-   Shah et al., U.S. patent application Ser. No. 10/919,518.-   Shah et al., U.S. patent application Ser. No. 10/932,295.-   Shah et al., U.S. patent application Ser. No. 10/972,089.-   Shah et al., U.S. patent application Ser. No. 11/153,435.-   Dake et al., U.S. patent application Ser. No. 14/210,284.-   Dake et al., U.S. patent application Ser. No. 14/698,150.-   Dake et al., U.S. patent application Ser. No. 14/698,274.-   Dake et al., U.S. patent application Ser. No. 14/698,332.-   Dake et al., U.S. patent application Ser. No. 14/850,405.-   USDA SR-21 released Dec. 7, 2011 by U.S. Department of Agriculture.

We claim:
 1. A method of preparing a composition, comprising: (a)applying a starting material to a filtration means, said filtrationmeans having a pore size of 500 nanometers (nm) or smaller, saidstarting material being prepared from an animal source, (b) allowing thestarting composition to pass through the filtration means, and (c)collecting retentate that is retained on said filtration means to obtainsaid composition, wherein said composition obtained in step (c)comprises more than 3% hydroxyproline by weight of total amino acids insaid composition.
 2. The method of claim 1, wherein said filtrationmeans is selected from the group consisting of microfiltration,ultrafiltration, nanofiltration, reverse osmosis, and combinationthereof.
 3. The method of claim 1, wherein the starting material isobtained by extracting proteins from poultry parts.
 4. The method ofclaim 3, wherein said poultry parts comprise 50-100% (w/w) of poultrybones and cartilage.
 5. The method of claim 3, wherein said poultryparts comprises 50-100% (w/w) of poultry cartilage.
 6. The method ofclaim 1, wherein said retentate obtained in step (c) comprises more than50% (w/w) collagen.
 7. The method of claim 6, wherein said collagencomprises more than 50% (w/w) Type II collagen.
 8. The method of claim1, wherein no enzyme is added to said starting material before beingapplied to the filtration means.
 9. The method of claim 3, furthercomprising a cooking step prior to step (a), wherein the poultry partsare cooked in water at a temperature 250 F or higher for 6 hours orlonger during the cooking step.
 10. The method of claim 1, wherein saidfiltration means has a pore size of 100 nm or smaller.
 11. The method ofclaim 1, wherein said filtration means has a pore size of 50 nm orsmaller.
 12. The method of claim 1, wherein said composition obtained instep (c) comprises more than 4% hydroxyproline by weight of total aminoacids in said composition.
 13. The method of claim 1, further comprisinga step of collecting permeate that passes through said filtration meansto obtain a pumpable composition.
 14. The method of claim 1, whereinsaid starting material is subjected to a separation step before beingapplied to the filtration means, said separation step preceding step(a).
 15. The method of claim 14, wherein said separation step is carriedout by centrifugation.